The Mexican ground squirrel (Spermophilus mexicanus) has a natural resistance to western diamondback rattlesnake venom (Crotalus atrox). The LD50 for the Mexican ground squirrel is 53 mg/kg body weight, which is 13 times higher than that of BALB/c mice. An antihemorrhagic factor from serum of the Mexican ground squirrel was isolated using Sephadex G-200 gel filtration, ion exchange A-50, G-75 gel filtration and HPLC DEAE 5PW ion exchange chromatography. The purified factor neutralized proteolytic and hemorrhagic activity of crude C. atrox venom. The results of this research suggest that the antihemorrhagic factor in the serum of the Mexican ground squirrel is not an antibody and neutralizes hemorrhagic activity of C. atrox venom.